The objective of this proposal is to develop a novel technology for detecting multiple protein markers for disease management. In this project, the technology will be used in profiling the phosphorylation state of numerous proteins in a single test. Most clinical tests currently in use are based on the detection of single disease markers, which limits their accuracy. In contrast, a single test to evaluate multiple disease markers would significantly enhance the analytical and predictive powers of disease testing. Therefore, developing a high throughput and cost effective technology for multi-marker detection is an urgent, unmet, medical need. Recent research suggests that certain protein phosphorylation events are likely to occur during specific diseased states. This program will develop a high throughput protein detection technology, Epitope-based Quantification Using Antibodies and T7-polymerase (EpiQUANT), for determining the phosphorylation patterns of numerous proteins at once in normal and diseased tissue samples. The diagnostic potential of EpiQUANT technology will be evaluated using clinical samples. Until now, most phospho-specific antibodies have been polyclonal in nature, which severely limits their diagnostic utility. The advent of Epitomics' rabbit monoclonal antibody (RabMAb) technology enables, for the first time, the reliable production of monoclonal phospho-specific antibodies with consistent quality and an unlimited supply. EpiQUANT will represent a new level of throughput through the novel, synergistic combination of two proven technologies-antibody binding and DNA microarray detection. If successfully developed, EpiQUANT technology and diagnostic kits will enable the simultaneous, systematic analysis of phosphorylated proteins and relate these patterns directly to diseased versus healthy states. These reagent kits will be commercialized to the research community and further developed into diagnostic products.